Growing Mycobacterium

[Irka]

In article <goldberg-1703971007070001 at 140.176.74.203>, goldberg at bms.com
(Steven Goldberg) wrote:

> I am trying to grow Mycobacterium vaccae obtained from ATCC.  This is a
> methylotrophic bacterium, so I have tried various minimal and rich media
> for Mycobacterium with and without methanol.  So far (after 3 days at 37
> deg. C) I only see faint growth.  ATCC recommends using Lowenstein-Jensen
> medium, but this calls for a solution of hen's eggs, among other strange
> ingredients which I do not possess.  Does anyone have a simpler medium
> that I could try for liquid growth?  Since I have a pure culture, I do not
> need to differentiate or isolate colonies (which is what the
> Lowenstein-Jensen medium was intended to do, as far as I can tell).
> 
> Thanks for any assistance.
> 
> 
> Steve

Steve:

I grow Mycobacterium phlei for our student laboratories in brain heart
infusion (BHI) broth/agar medium supplemented with glycerol (I'm not at
work, so I can't give you the glycerol concentration right now).  I find
that, unless you are inoculating from a fresh culture, it takes at least 3
days, usually 5 days, for any visible signs of growth, and 7 days for good
growth.  I would suggest incubating your culture for at least 5 to 7 days
before you give up on any particular medium that you are trying.

I don't know if BHI with glycerol, perhaps supplemented also with methanol,
would be an appropriate medium for your strain, but you could give it a
try.  I can send you further information on the exact concentration of
glycerol, if you are interested.

-- 
Irka